Radiation damage of protein crystals at cryogenic temperatures between 40 K and 150 K

Teng, Tsu-Yi; Moffat, Keith

doi:10.1107/s0909049502008579

Cited by 58 publications

(58 citation statements)

References 14 publications

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“…Using the radiationdamage metric of Kmetko et al (2006), we examined 49 thaumatin crystals to determine the radiation-sensitivity at 11 temperatures from 300 to 100 K. Our results agree with previous studies at 300 and 100 K and indicate that most of the reduction in sensitivity on cooling from 300 to 100 K occurs above 200 K. These results are consistent with previous reports on the temperature-dependence of global damage (Teng & Moffat, 2002;Borek et al, 2007;Meents et al, 2010). We model radiation damage as a thermally activated process with a large barrier and a small barrier.…”

Section: Radiation-damage Mechanismssupporting

confidence: 82%

Glass transition in thaumatin crystals revealed through temperature-dependent radiation-sensitivity measurements

Warkentin

Thorne

2010

Acta Crystallogr D Biol Crystallogr

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The temperature-dependence of radiation damage to thaumatin crystals between T = 300 and 100 K is reported. The amount of damage for a given dose decreases sharply as the temperature decreases from 300 to 220 K and then decreases more gradually on further cooling below the protein-solvent glass transition. Two regimes of temperature-activated behavior were observed. At temperatures above $200 K the activation energy of 18.0 kJ mol À1 indicates that radiation damage is dominated by diffusive motions in the protein and solvent. At temperatures below $200 K the activation energy is only 1.00 kJ mol À1 , which is of the order of the thermal energy. Similar activation energies describe the temperaturedependence of radiation damage to a variety of solvent-free small-molecule organic crystals over the temperature range T = 300-80 K. It is suggested that radiation damage in this regime is vibrationally assisted and that the freezing-out of amino-acid scale vibrations contributes to the very weak temperature-dependence of radiation damage below $80 K. Analysis using the radiation-damage model of Blake and Phillips [Blake & Phillips (1962), Biological Effects of Ionizing Radiation at the Molecular Level, indicates that large-scale conformational and molecular motions are frozen out below T = 200 K but become increasingly prevalent and make an increasing contribution to damage at higher temperatures. Possible alternative mechanisms for radiation damage involving the formation of hydrogen-gas bubbles are discussed and discounted. These results have implications for mechanistic studies of proteins and for studies of the protein glass transition. They also suggest that data collection at T ' 220 K may provide a viable alternative for structure determination when cooling-induced disorder at T = 100 is excessive.

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Section: Radiation-damage Mechanismssupporting

confidence: 82%

Glass transition in thaumatin crystals revealed through temperature-dependent radiation-sensitivity measurements

Warkentin

Thorne

2010

Acta Crystallogr D Biol Crystallogr

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“…The former results from the interaction of an X-ray photon with atoms in the sample, leading to the ejection of a highly energetic electron as a result of the photoelectric effect, which is the dominant inelastic event at the photon energies used in macromolecular crystallography (Murray et al, 2005). Primary radiation damage is temperature-independent (Teng & Moffat, 2002). Secondary damage arises from the many secondary radicals created by the primary photoelectron.…”

Section: Protein Structures At Various Cryo-temperaturesmentioning

confidence: 99%

Temperature-dependent macromolecular X-ray crystallography

Weik

Colletier

2010

Acta Crystallogr D Biol Crystallogr

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X-ray crystallography provides structural details of biological macromolecules. Whereas routine data are collected close to 100 K in order to mitigate radiation damage, more exotic temperature-controlled experiments in a broader temperature range from 15 K to room temperature can provide both dynamical and structural insights. Here, the dynamical behaviour of crystalline macromolecules and their surrounding solvent as a function of cryo-temperature is reviewed. Experimental strategies of kinetic crystallography are discussed that have allowed the generation and trapping of macromolecular intermediate states by combining reaction initiation in the crystalline state with appropriate temperature profiles. A particular focus is on recruiting X-ray-induced changes for reaction initiation, thus unveiling useful aspects of radiation damage, which otherwise has to be minimized in macromolecular crystallography.

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“…In an important advance, it was shown that radiation damage could be mitigated by cryogenic vitrification of protein crystals (Hope 1988). At temperatures of 100 K, crystals become almost (but not quite) immortal (Garman & Schneider 1997;Teng & Moffat 2002;Meents et al 2010). Control studies with small proteins showed that in general cryo-cooling changes neither the overall structure nor details of the functional sites but that the dynamic properties of a protein may be changed (Rasmussen et al 1992).…”

Section: Radiation Damage and Cryo-crystallographymentioning

confidence: 99%

Macromolecular crystallography at synchrotron radiation sources: current status and future developments

Duke

Johnson

2010

Proc. R. Soc. A.

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X-ray diffraction with synchrotron radiation (SR) has revealed the atomic structures of numerous biological macromolecules including proteins and protein complexes, nucleic acids and their protein complexes, viruses, membrane proteins and drug targets. The bright SR X-ray beam with its small divergence has made the study of weakly diffracting crystals of large biological molecules possible. The ability to tune the wavelength of the SR beam to the absorption edge of certain elements has allowed anomalous scattering to be exploited for phase determination. We review the developments at synchrotron sources and beamlines from the early days to the present time, and discuss the significance of the results in providing a deeper understanding of the biological function, the design of new therapeutic molecules and time-resolved studies of dynamic events using pump-probe techniques. Radiation damage, a problem with bright X-ray sources, has been partially alleviated by collecting data at low temperature (100 K) but work is ongoing. In the most recent development, free electron laser sources can offer a peak brightness of hard X-rays approximately 10 8 times brighter than that achieved at SR sources. We describe briefly how early experiments at FLASH and Linear Coherent Light Source have shown exciting possibilities for the future.

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Radiation damage of protein crystals at cryogenic temperatures between 40 K and 150 K

Cited by 58 publications

References 14 publications

Glass transition in thaumatin crystals revealed through temperature-dependent radiation-sensitivity measurements

Glass transition in thaumatin crystals revealed through temperature-dependent radiation-sensitivity measurements

Temperature-dependent macromolecular X-ray crystallography

Macromolecular crystallography at synchrotron radiation sources: current status and future developments

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